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EP-4735892-A1 - NON-INVASIVE DIAGNOSTIC BIOMARKER-BASED TEST OF ENDOMETRIOSIS

EP4735892A1EP 4735892 A1EP4735892 A1EP 4735892A1EP-4735892-A1

Abstract

The present invention relates to an in vitro method for assessing whether a subject has endometriosis comprising a step of measuring, in a biological sample of the subject, the expression level of at least two biomarkers, wherein the at least two biomarkers are MUC-16 and either ANXA1 or ARHGAP1.

Inventors

  • BERTHOUMIEUX, Sara
  • MIGNON-GODEFROY, Karine
  • Real, Cécile

Assignees

  • Endodiag

Dates

Publication Date
20260506
Application Date
20240628

Claims (15)

  1. 1. An in vitro method for assessing whether a subject has endometriosis comprising a step of measuring, in a biological sample of the subject, the expression level of at least two biomarkers, wherein the at least two biomarkers are MUC-16 and either ANXA1 or ARHGAP1.
  2. 2. The in vitro method according to claim 1, wherein the method comprises measuring the expression level of at least three biomarkers, wherein the at least three biomarkers are: • MUC-16, and • ANXA1 or ARHGAP 1 , and • at least one biomarker selected from the group comprising or consisting of EREG, ANXA1, ARHGAP1, t-PA, PCSK9, ARSB, ESM-1 and TMPRSS15.
  3. 3. The in vitro method according to claim 1 or claim 2, wherein the method comprises measuring the expression levels of at least three biomarkers, wherein the at least three biomarkers are: • MUC- 16, ANXA1 and EREG, or • MUC-16, ANXA1 and ARSB, or • MUC-16, ANXA1 and TMPRSS15, or • MUC- 16, ARHGAP 1 and PC SK9, or • MUC- 16, ARHGAP 1 and t-PA.
  4. 4. The in vitro method according to claim 1 or claim 2, wherein the method comprises measuring the expression levels of at least five biomarkers, wherein the at least five biomarkers are MUC-16, ANXA1, EREG, t-PA and ESM-1.
  5. 5. The in vitro method according to any one of claims 1 to 4, wherein the method comprises determining a score based on the combination of the measured expression levels using a linear model.
  6. 6. The in vitro method according to any one of claims 1 to 4, wherein the method comprises determining a score based on the combination of the measured expression levels using a complex model.
  7. 7. The in vitro method according to any one of claims 1 to 6, wherein the biological sample is selected from the group consisting of any tissue, cell, fluid, saliva, tears, urine, sweat, sputum, liquid biopsy, blood, serum, plasma, ascites, cyst fluid, vaginal fluid and cervico-vaginal fluid, preferably the biological sample is blood, serum or plasma.
  8. 8. The in vitro method according to any one of claims 1 to 7, wherein the level of the biomarkers is measured by immunoassays, including ELISA, Western blot, immune-electrophoresis, immunostaining and/or Proximity Extension Assay (PEA) ; enzymatic activity ; flow cytometry ; spectrometry, including mass spectrometry ; RT-PCR ; RT-qPCR ; Northern Blot ; or hybridization techniques.
  9. 9. A diagnostic device, comprising means for measuring the expression level of biomarkers of a signature comprising at least two biomarkers, wherein the at least two biomarkers are MUC-16 and either ANXA1 or ARHGAP1.
  10. 10. The diagnostic device according to claim 9, wherein the signatures comprises at least three biomarkers, wherein the at least three biomarkers are: • MUC- 16, ANXA1 and EREG, or • MUC-16, ANXA1 and ARSB, or • MUC-16, ANXA1 and TMPRSS15, or • MUC- 16, ARHGAP 1 and PC SK9, or • MUC- 16, ARHGAP 1 and t-P A.
  11. 11. The diagnostic device according to claim 9 or claim 10, wherein the signature comprises at least five biomarkers, wherein the at least five biomarkers are MUC- 16, ANXA1, EREG, t-PA and ESM-1.
  12. 12. The diagnostic device according to any one of claims 9 to 11, comprising i) capture reagents specific for the biomarkers of the signature, and ii) detection reagents for detecting the biomarkers of the signature.
  13. 13. The diagnostic device according to claim 12, wherein the capture reagents are bound to plates, chips, beads, membranes or arrays.
  14. 14. The diagnostic device according to claim 12 or claim 13, wherein the detection reagents are enzymes, radioactive proteins, or fluorescent proteins.
  15. 15. The diagnostic device according to any one of claims 12 to 14, wherein the means for measuring the expression level of biomarkers of a signature comprise or consist of means that may be used in immunoassays, including ELISA, Western blot, immune-electrophoresis, immunostaining or Proximity Extension Assay (PEA) ; enzymatic activity ; flow cytometry ; spectrometry, including mass spectrometry ; RT-PCR ; RT-qPCR ; Northern Blot ; or hybridization techniques.

Description

NON-INVASIVE DIAGNOSTIC BIOMARKER-BASED TEST OF ENDOMETRIOSIS FIELD OF INVENTION [0001] The present invention relates to an in vitro non-invasive diagnostic biomarkerbased test of endometriosis and its application. BACKGROUND OF INVENTION [0002] Endometriosis is defined as the presence of endometrial-like tissue outside the uterine cavity. This chronical disease is considered a debilitating gynecological pathology with a high incidence, between 6-10% among women of childbearing age. The prevalence of endometriosis has been estimated at 180 million women worldwide. Endometriosis lesions can occur at different locations in the body, including the pelvic peritoneum or ovary or can infiltrate pelvic structures below the peritoneal surface, thus defining three clinical phenotypes, superficial endometriosis (SUP), endometrioma (OMA) and deep endometriosis (DIE). The main symptoms consist of chronic pelvic pain, dysmenorrhea, dyspareunia, and infertility. Given the nonspecific symptoms of this disease, the gold standard for a definitive diagnosis is based on surgical procedure such as laparoscopy to visualize the lesions followed by a histological confirmation. The average delay between the onset of symptoms and diagnosis is 7 years. The lack of specific and heterogenous symptoms associated with a stigma or symptom normalization, the lack of awareness of the pathology by the general public, and by many practitioners and some gynecologists, are in part responsible for the diagnostic delay. [0003] The management of the patient is evolving, and no systematic surgery and excision of the lesions are nowadays recommended. In fact, according to several gynecological scientific societies, there is a consensus to consider the patient’s predominant symptoms and/or the desire for a child before considering invasive treatment. Available medical therapies for endometriosis include non-hormonal treatments such as painkillers and hormonal treatments such as combined oral contraceptives, progestins and gonadotropin-releasing hormone analogues. [0004] Medical imaging has led to substantial improvements in the diagnosis of endometriosis. Transvaginal ultrasonography (TVUS) and / or magnetic resonance imaging (MRI) can be used to establish a diagnosis of OMA and DIE. However, SUP can usually not be visualized by imaging since the size and location of the lesions is below the threshold for detection. In addition, few radiologists are aware of the specific imaging modalities for the disease and patients who are suspected of endometriosis should be referred to a specialist of this disease. [0005] In a clinical practice, a noninvasive diagnostic biomarker-based test could help shorten the time during which the patient does not know what she is suffering from and is not taken care of. The identification of validated biomarkers in blood is an unmet medical need that demands to be fulfilled. Many publications and patents are available describing the use of biomarkers alone (miRNA, protein, metabolites, RNA, circulating free DNA, antibodies, etc.) and/or in combination with various clinical performances. [0006] W02021/168040 discloses compositions and methods that provide a high degree of sensitivity and a high degree of specificity for the pre-operative assessment of endometriosis in pre-menopausal women having a variety of endometriosis types. W02021/168040 discloses a large panel of more than 50 polypeptide biomarkers that are allegedly differentially present in subjects having endometriosis. [0007] However, there is still a need for improved noninvasive diagnostic methods that not only have a high degree of sensitivity, but that also provide a high degree of specificity for diagnosing endometriosis and allowing a further adapted treatment, based on a small number of biomarkers to be measured, such as, for example, based on as few as five biomarkers. SUMMARY [0008] This invention thus relates to an in vitro method for assessing whether a subject has endometriosis comprising a step of measuring, in a biological sample of the subject, the expression level of at least two biomarkers, wherein the at least two biomarkers are MUC-16 and either ANXA1 or ARHGAP 1. [0009] In some embodiments, the method comprises measuring the expression level of at least three biomarkers, wherein the at least three biomarkers are: • MUC-16, and • ANXA1 or ARHGAP 1 , and • at least one biomarker selected from the group comprising or consisting of EREG, ANXA1, ARHGAP1, t-PA, PCSK9, ARSB, ESM-1 and TMPRSS15. [0010] In some embodiments, the method comprises measuring the expression levels of three biomarkers, wherein the three biomarkers are: • MUC- 16, ANXA1 and EREG, or • MUC-16, ANXA1 and ARSB, or • MUC-16, ANXA1 and TMPRSS15, or • MUC- 16, ARHGAP 1 and PC SK9, or • MUC- 16, ARHGAP 1 and t-PA. [0011] In some embodiments, the method comprises measuring the expression levels of at least three biomarkers, wherein the at least three biomarkers are: • MUC- 16, ANX